OBJECTIVES: To investigate the ethnic origin of Chuanqing people, one of the largest unidentified ethnic groups in Guizhou, China, and analyze its genetic relationships with surrounding populations. METHODS: Based on a self-developed microhaplotype system, we conducted genotyping and analyzed the genetic distribution of microhaplotype loci and their forensic applicability in Chuanqing population in Guizhou Province. Using the microhaplotype data from different intercontinental populations and previously reported data from Han population living in Guizhou Province, we systematically investigated the genetic background of Chuanqing people through population genetic approaches, including genetic distance estimation, principal component analysis, and phylogenetic tree construction. RESULTS: Among the studied population, the number of haplotype per microhaplotype ranged from 6 to 25. The average expected heterozygosity (He), observed heterozygosity (Ho), power of discrimination (PD), and probability of exclusion (PE) were 0.8291, 0.8301, 0.9387, and 0.6593, respectively. The cumulative power of discrimination (CPD) and cumulative probability of exclusion (CPE) for these 33 loci were 1-2.62×10^-41 and 1-7.64×10^-17, respectively. Population genetic analyses revealed that the Chuanqing population had close genetic relationships with the East Asian populations, especially the local Guizhou Han population, Beijing Han population and the Han populations living in southern China. CONCLUSIONS The 33 microhaplotypes exhibit high levels of genetic diversity in the Guizhou Chuanqing population, highlighting their potentials for both forensic identification and parentage testing. The Han populations might have contributed a significant amount of genetic material to the Chuanqing population during the formation and development of the latter.
Humans ; China/ethnology* ; Ethnicity/genetics* ; Forensic Genetics/methods* ; Genetics, Population ; Genotype ; Haplotypes ; Phylogeny ; East Asian People/genetics*

OBJECTIVE To establish a method for determining the content of 11 antioxidants and their degradation products in recombinant Exendin-4-FC fusion protein injection by HPLC. METHODS The protein was precipitated with saturated ammonium sulfate. After centrifugation, the supernatant was transferred to a C18 solid phase extraction cartridge activated by methanol. Then the cartridge was eluted with 4 mL of methanol and 5 mL of ethyl acetate respectively, and the eluent was diluted with methanol-ethyl acetate(2∶3) mixed solvent and passed through a 0.22 µm PTFE hydrophobic filter. It was analyzed by HPLC and quantified by external standard method. Chromatographic conditions: Kinetex® XB-C18 100Å (100 mm×4.6 mm, 2.6 µm)column, the detection wavelength was 230 nm, the column oven was 30 ℃, the injection volume was 5 µL and the flow rate was 0.4 mL·min–1, mobile phase was 0.1% formic acid-methanol(A)-0.1% formic acid aqueous solution(B), the running time was 45 min. RESULTS The 11 target substances showed a good linear relationship in the range of 2.5−35 μg·mL–1 with R2 ≥0.99. At three different concentration(25, 10, 5 μg·mL–1) of spiked samples, the average recovery rates of 11 antioxidants ranged from 88.1% to 106.5%, with RSDs in the range of 0.10%–9.05%. The RSDs of 6 repeatable samples was 2.01%–4.77%, which of 12 intermediate precision samples was 2.58%–9.75%. The positive/inverted samples of three batches of recombinant Exendin-4-FC fusion protein injection were detected at 0 month, 3 months and 6 months(25 ℃), and the results showed that there was no antioxidant and its degradation leaching in all batches of samples at different detection points. CONCLUSION The method has good specificity, high accuracy and precision, good solution stability, high durability and can be used for the content detection of antioxidants in drugs.

Objective:To construct a plasmid for expression Mycobacterium tuberculosis ( Mtb) Rv3133c and to evaluate the immunogenicity of Rv3133c through population and mice experiments. Methods:The recombinant expression plasmid pPROEX-Rv3133c was constructed. The transformed E. coli BL21 (DE3) carrying expression plasmid was induced by IPTG to express the recombinant Rv3133c (rRv3133c). Western blot was used to identify the expressed protein. Whole-blood IFN-γ release assay (WBIA) was preformed to assess the immunogenicity of rRv3133c in Mtb-infected population. Antigen-specific antibodies in serum, Th1 type cytokines in splenocytes, functional T cell subset responses in splenocytes and the expression of cytokines at mRNA level in lung tissues were detected after immunizing mice subcutaneously with rRv3133c and adjuvant DC. Results:The rRv3133c was constructed and expressed successfully. Stimulation with rRv3133c promoted the production of IFN-γ in Mtb-infected population, especially in patients with latent tuberculosis infections. After immunizing mice with rRv3133c+ DC, the levels of IFN-γ, TNF-α and IL-2, the number of IFN-γ + TNF-α + CD4 + T cells in spleen and the expression of antigen-specific IFN-γ, TNF-α and iNOS at mRNA level in lung tissues were higher than those in BCG-immunized mice, but lower than those in BCG+ rRv3133c+ DC group. The serum IgG2a/IgG1 ratios in the rRv3133c+ DC group and the BCG+ rRv3133c+ DC group were greater than 1, and significantly higher than that of the BCG group. Conclusions:The rRv3133c had good immunogenicity and could induce strong Th1 immune response, suggesting that it was a potential candidate antigen for subunit vaccine against tuberculosis.

Objective:To evaluate the effects of a booster immunization with a candidate tetanus toxoid, reduced diphtheria toxoid and acellular pertussis combined vaccine (Tdap) in a rat model after primary vaccination with diphtheria, tetanus, acellular pertussis and Sabin strain inactivated poliovirus combined vaccine (DTacP-sIPV) or diphtheria, tetanus, acellular pertussis, inactivated poliovirus and haemophilus type b combined vaccine (DTacP-IPV/Hib) for further preclinical study.Methods:Wistar rats were randomly divided into three groups and respectively immunized with a self-developed DTacP-sIPV, a marketed DTacP-IPV/Hib and normal saline at 0, 1, and 2 months of age. Serum levels of antibody against each component in each group were detected before immunization and after each dose. A booster dose of the candidate Tdap was given 10 months after primary immunization. Serum levels of antibody against each component in each group were detected before, 1 month and 6 months after the booster immunization.Results:One month after three doses of primary immunization, the geometric mean titers (GMT, Log2) of antibodies against diphtheria toxoid (DT), tetanus toxoid (TT), pertussis toxin (PT), filamentous hemagglutinin (FHA) and pertactin (PRN) in the DTacP-sIPV group were 17.41, 18.34, 18.11, 19.93 and 13.91, respectively, and the seroconversion rates of these components all reached 100%. Ten months after primary immunization, the GMTs of antibodies against DT, TT, PT, FHA and PRN decreased to 15.17, 14.26, 13.60, 14.51 and 10.39, respectively, and the seroconversion rates remained above 89%. One month after booster immunization, the GMTs of antibodies against DT, TT, PT and FHA in the DTacP-sIPV and DTacP-IPV/Hib groups were 16.49/17.26, 16.80/17.63, 16.70/17.74 and 18.48/19.26, respectively, and the seroconversion rates of these components all reached 100% with no significant difference between the two groups ( P>0.05). The GMTs of anti-PRN antibody in the DTacP-sIPV and DTacP-IPV/Hib groups were 13.07 and 11.00, and the seroconversion rates were 100% and 88%, which were higher in the DTacP-sIPV group than in the DTacP-IPV/Hib group ( P<0.05). Six months after booster immunization, the GMTs of antibodies against DT, TT, PT, FHA and PRN in the DTacP-sIPV and DTacP-IPV/Hib groups decreased to 15.74/14.87, 15.07/15.14, 14.84/15.73, 16.62/16.37 and 11.44/9.96, respectively, and the seroconversion rates remained above 88%. Conclusions:Booster vaccination with the candidate Tdap vaccine induces humoral immune response following primary immunization with DTacP-sIPV or DTacP-IPV/Hib in the Wistar rat model, while the antibody titer decreases with time.

Objective To develop urine AD7C-NTP diagnostic kit,analyze and evaluate its application value on AD.Methods Immunogenicity AD7C-NTP peptide fragments had synthesized by solidphase methods.The animals immunized to prepare antibodies.After matching screening.mouse antibody was uesed as coating antibody.biotin-labeled rabbit antibody wag used as testing antibody,and horseradish peroxidase was labeied with avidin.The urine AD7C-NTP ELISA detective method was established.The AD7C-NTP levels in morning urine samples of 121 AD patients and 118 age-matched controls were collected.Results AD7C-NTP antibodies were identified.Mouse anti-AD7C-NTP antibody titer in ELISAwas 1:8 000,and rabbit anti-AD7C-NTP antibody titer in ELISA was 1:32 000:WB was uesd to detect human brain specimens and there was a single band with molecular weight of 41 000.The lowst detection limit of ELISA methodology was 0.5μg/L The linear range was 0-10μg/L,normal reference value ≤1.5μg/J,the average recovery rate was 100.2%.The intra and inter of CV were 3.8%,4.5%,7.6%,6.8% respectively.The AD7C-NTP levels[2.25(0.43-8.62)μg/L]of urine in AD group was higher than those in contorl group[0.82(0.47-2.77)μg/L,P<0.01].The positive rates in AD group and control group were 89.3% and 15.3% respectively.The sensitivity Was 89.3%and specificity was 84.7%.Conclusions The animals are immunized with the self-designed synthetic peptide fragment to prepare AD7C-NTP antibodies successfully.The established ELISA method for detection of urine AD7C-NTP with high sensitivity,and precision can be used as an assistant examination in clinical diagnosis of AD.

Objective To establish an ultraviolet-irradiation damage model in cultured fibroblasts derived from human skin and to explore the potential protective effects and mechanisms of amyloid precursor protein 17-met peptide (APP17-mer peptide) on the oxidative damage and collagen metabolism in cultured fibroblasts after ultraviolet irradiation. Methods Human dermal fibroblast cultures were established by outgrowth from foreskin biopsies of a healthy donor and were irradiated by a single exposure to ultraviolet rays and cultured in a series of concentrations of APP17-mer peptide (0, 20, 40, 80 μmol/L).The activity of fibroblasts was detected by the assay of MTT. The intracellular ROS level was measured with a confocal microscope. The expression of MMP-1 mRNA was analyzed real-time quantitatively following RT-PCR. Results Primary cultures of human skin fibroblasts were established from human foreskin in DMEM supplemented with 10 % fetal bovine serum. UV irradiation depressed cellular activity and increased intracellular level of ROS (P<0.05). 40μmol/L and 80μmol/L APP17-mer peptide increased the cellular activity in both UV irradiated fibroblasts and unirradiated fibroblasts (P<0.05), however,20 μmol/L did not show such protective effects (P>0. 05). 40μmol/L APP17-mer peptide could depress the level of ROS in irradiated libroblasts. A single exposure of fibroblasts to UV irradiation resulted in 1.78 foldup-regulation of MMP-1 mRNA compared with unirradiated sample, 40μmol/L and 80μmol/L APP17-mer peptide decreased the expression of MMP-1 mRNA (P<0.05 and P<0.01, respectively).Conclusion APP17-mer peptide can enhance cellular activity under UV-induced oxidative stress and in-hibit collagen degradation in fibroblasts irradiated with ultraviolet rays. Inhibition of ROS production may be involved in the protective mechanism of APP17 peptide.

Objective To study the CT features of nasal bone foramen and nasal bone fracture,and investigate the differential key points of them.Methods visual observation was performed for 30 selected skull specimens of adults(60 nasal bones)(specimens group),then CT imaging was obtained for them with volume redering(VR)reconstruction(imaging group).The frequency of the nasal bone foramens,their size,shape and location were statistical analysed.Ferthermore,CT features of nasal bone foramen and nasal bone fracture were analyzed in 30 patients who were suspected having nasal bone fractures by clinicians.Results Forty foramen shape nasal bone foramens were found in observation group,while 37 were obtained in imaging group.For patients,19 cases were diagnosed as single fracture of nasal bone by non-spiral axial high-resolution CT images.When using both axial and multi-plane reformation(MPR)and VR images,8 of 19 cases were diagnosed as single fracture,5 were multiple fractures,4 were normal nasal bone foramen,and 2 were normal variation.The other 11 cases that were diagnosed as multiple fractures by non-spiral axial HRCT images were demonstrated as 6 cases multiple fractures and 5 single fracture by combining axial images with MPR and VR images.The frequencies of foramen shape nasal bone foramen in specimens group and imaging group were 66.7% and 61.7%,respectively,and there were no significant differences between them(χ2=1.33,P＞0.05).Conclusion The nasal bone foramen is the normal anatomy,and to recognize the imaging findings of it can improve the diagnostic accuracy of nasal bone fracture.

Objective To evaluate the safety of Fotemustine. Methods The studies had been done according to the experimental method of injection agents in pharmacodynamics guide principle on toxic test methods of application. The vascular stimulative assay of fotemustine injection on rabbit, the hemolyzation or agglutination of it in vitro, and the allergic reaction on guinea pig were performed. Results It was showed that the Fotemustine injection had no vascular irritation and injury effect on rabbit vascular endothelial cell. Compared with that of the control group, there was no difference. Conclusion No evidences of hemolyzation and aggregation on rabbit erythrocyte, and no allergic reaction on guinea pig were observed.

Urinary microquantitative albumin (UALb), immunoglobulin G (UIgG) ?2-microglo-bulin (U?2m), and Tamm-Horsfall protein (THP) were measured in 36 chronic B hepatitis (CHB) and 23 liver cirrhosis (LC) patients with radioimmunoassay. 109 healthy normal subjects served as controls. The results showed that the excretion of UALb was significantly increased in 17 patients. Out of 17 patients,the excretion of UIgG in 5 and U?2m in 16 were increased and the excretion of THP in 9 patients were decreased. Compared with normal controls there was a significant difference (P